Molecular analysis of the Penicillium marneffei glyceraldehyde-3-phosphate dehydrogenase-encoding gene (gpdA) and differential expression of gpdA and the isocitrate lyase-encoding gene (acuD) upon internalization by murine macrophages.
Penicillium marneffei is an intracellular dimorphic fungus that can cause a fatal disseminated disease in human immunodeficiency virus-infected patients. The factors that affect the pathogenicity of this fungus remain unclear. Here, we report the isolation and characterization of the gpdA cDNA and genomic clones encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in P. marneffei. Phylogenetic analysis of GAPDH amino acid sequences demonstrated the evolutionary relationship of P. marneffei to other fungi, including the intracellular pathogen Ajellomyces capsulatus. To assess the central importance of phagocytic cells in defence against P. marneffei infection, we used Northern blotting to investigate the response of the isocitrate lyase-encoding gene (acuD) and gpdA to nutrient deprivation inside macrophages. The results revealed that after macrophage internalization, the gene involved in the glyoxylate cycle, acuD, showed higher expression levels as early as 2 h from the start of co-incubation, and the differential expression could be observed again at 8 h after infection. In contrast, the expression of gpdA was downregulated in the yeast phase, as well as during macrophage infection after 2, 4 and 8 h of infection. The induction of P. marneffei acuD was shown to be coordinated with the downregulation of the glycolytic gpdA gene, implying that the cytoplasmic environment of macrophages is deficient in glucose and the glyoxylate pathway could be used by this pathogen to allow subsistence on two-carbon compounds within the host cell following its intracellular persistence.
Thirach
Sophit
S
Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
Cooper
Chester R
CR
Department of Biological Sciences, Youngstown State University, Youngstown, OH, USA.
Vanittanakom
Nongnuch
N
Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D007522:Isocitrate Lyase / Q000235:genetics*; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D008969:Molecular Sequence Data; D010407:Penicillium / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny
D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D007522:Isocitrate Lyase